Cotransport of H+, lactate and H2O by membrane proteins in retinal pigment epithelium of bullfrog.

1. The interaction between H+, lactate and H2O fluxes in the retinal membrane of the pigment epithelium from bullfrog Rana catesbiana was studied by means of ion-selective micro-electrodes. 2. Changes in intracellular pH and cell volume were recorded in response to abrupt changes in retinal solution concentration and/or osmolarity. 3. Two parallel pathways for water transport were identified across the retinal membrane, an osmotic one with a hydraulic water permeability of 3.2 x 10(-4) cm s-1 (osmol l-1)-1 and one which depended on the presence of lactate. 4. Addition of sodium lactate to the retinal solution caused cell shrinkages that were small compared with those produced by mannitol. The reflection coefficient for sodium lactate was 0.25. 5. Isosmotic replacement of Cl- with lactate caused an influx of water. Simultaneous acidification of the retinal solution from pH 7.4 to 6.4 enhanced the effect. The influx of water could proceed against osmotic gradients elicited by mannitol. 6. The interdependence of the fluxes of H+, lactate and H2O can be described as cotransport: the fluxes had a fixed ratio of about 109 mmol of lactic acid per litre of water, the flux of one species was able to energize the flux of the other two, and the fluxes exhibited saturation for increasing driving forces. 7. The Gibbs equation gives an accurate quantitative description of these coupled fluxes.